Fig. 2.
Identification of α6 subunits in cerebellar membrane extracts. GABAA receptors were immunoprecipitated from cerebellar membrane extracts using α6(1–15) antibodies. Receptors were then subjected to SDS-PAGE and Western blot analysis, using digoxigenated antibodies.Lane 1, α6(429–434) antibodies;lane 2, α6(1–15) antibodies from rabbit 15; lane 3, α6(1–15) antibodies from rabbit 16. Antibodies bound to proteins were detected using anti-digoxigenin-alkaline phosphatase Fab fragments and a sensitive chemiluminescence detection system as described in Materials and Methods. The gel was calibrated with proteins of known molecular mass. The experiment was performed twice with similar results. The protein smear below the 56–57 kDa band was detected by all three antibodies and may have represented differentially glycosylated, partially degraded, or alternatively spliced α6 subunits.